| Summary: | Bartonellae are bacteria associated with mammals and their ectoparasites. Rodents often host different species of <i>Bartonella</i>. The aim of this study was to investigate the presence of <i>Bartonella</i> spp. in African giant pouched rats (<i>Cricetomys gambianus</i>) and their ectoparasites in Dakar, Senegal. In 2012, 20 rats were caught, and their fleas were identified. DNA was extracted from 170 selected fleas and qPCR was carried out to detect <i>Bartonella</i> spp. Subsequently, a <i>Bartonella</i> culture was performed from the rat blood samples and the isolated strains (<i>16S rRNA</i>, <i>rpoB</i>, <i>ftsZ</i> and ITS3) were genotyped. A total of 1117 fleas were collected from 19 rats and identified as <i>Xenopsylla cheopis</i>, the tropical rat flea. <i>Bartonella</i> DNA was detected in 148 of 170 selected fleas (87.1%). In addition, <i>Bartonella</i> strains were isolated from the blood of 17 rats (85%). According to <i>Bartonella</i> gene-sequence-based criteria for species definition, the isolated strains were identified as <i>B</i>. <i>massiliensis</i> (four strains) and two potential new species related to the zoonotic <i>B</i>. <i>elizabethae</i>. In this paper, these potentially new species are provisionally called <i>Candidatus</i> Bartonella militaris (11 strains) and <i>Candidatus</i> Bartonella affinis (two strains) until their description has been completed. <i>Cricetomys gambianus</i> and its fleas could constitute a public health risk in Dakar due to the high prevalence of <i>Bartonella</i> infection reported.
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